The effects of interleukin-6 in a rat model of hepatocellular carcinoma

Interleukin-6 (IL-6) is integrally linked to normal hepatic growth and liver regeneration. The aim of the present study was to determine the expression and function of IL-6 signalling pathways m a rat model of hepatocellular carcinoma (HCC). In these studies HCC was charactensed by dysregulated expression of IL-6 signalling components in vivo and in vitro, including downregulated expression of the IL-6 receptor and increased IL-6 production and secretion. Recombinant human IL-6 (rhIL-6) treatment of isolated rat HCC cells and hepatocytes in vitro led to the activation of janus kinase-signal transducer and activator of transcription (Jak-STAT) and Ras-mitogen activated protein kinase intracellular signalling pathways Cyclin dependent kinase (cdk) inhibitor expression was increased m HCC cells after treatment with IL-6, an effect that was not observed in isolated hepatocytes. Decreases in cdk activity after rhIL-6 treatment result in a G0/G1 growth arrest in HCC cells. As a result of these in vitro studies, an algmate based cell microencapsulation system was developed to provide continual in vivo rhIL-6 delivery in a rat model of HCC CHO cells transfected to supra-express rhIL-6 were encapsulated in algmate and implanted in HCC tumour burdened rats resulting in high serum levels of biologically active rhIL-6 Recombinant human IL-6 was detected in both normal and tumour liver tissues resulting in activation of the Jak-STAT pathway. Due to tumour volume variability, no conclusions on the effects of rhIL-6 on tumour progression in vivo could be determined However, rhIL-6 treatment did result in increased total hepatic weight. Collectively this study demonstrates a role for IL-6 m HCC progression and identifies algmate encapsulated CHO cells as an effective method for rhIL-6 delivery in vivo which has potential in the study of normal and abnormal liver growth

Puromycin-based vectors promote a ROS-dependent recruitment of PML to nuclear inclusions enriched with HSP70 and Proteasomes

<p>Abstract</p> <p>Background</p> <p>Promyelocytic Leukemia (PML) protein can interact with a multitude of cellular factors and has been implicated in the regulation of various processes, including protein sequestration, cell cycle regulation and DNA damage responses. Previous studies reported that misfolded proteins or proteins containing polyglutamine tracts form aggregates with PML, chaperones, and components of the proteasome, supporting a role for PML in misfolded protein degradation.</p> <p>Results</p> <p>In the current study, we have identified a reactive oxygen species (ROS) dependent aggregation of PML, small ubiquitin-like modifier 1 (SUMO-1), heat shock protein 70 (HSP70) and 20S proteasomes in human cell lines that have been transiently transfected with vectors expressing the puromycin resistance gene, puromycin n-acetyl transferase (pac). Immunofluorescent studies demonstrated that PML, SUMO-1, HSP70 and 20S proteasomes aggregated to form nuclear inclusions in multiple cell lines transfected with vectors expressing puromycin (puro) resistance in regions distinct from nucleoli. This effect does not occur in cells transfected with identical vectors expressing other antibiotic resistance genes or with vectors from which the pac sequence has been deleted. Furthermore, ROS scavengers were shown to ablate the effect of puro vectors on protein aggregation in transfected cells demonstrating a dependency of this effect on the redox state of transfected cells.</p> <p>Conclusion</p> <p>Taken together we propose that puromycin vectors may elicit an unexpected misfolded protein response, associated with the formation of nuclear aggresome like structures in human cell lines. This effect has broad implications for cellular behavior and experimental design.</p

"There is people like us and there is people like them, and we are not like them." Understating social exclusion - a qualitative study

Social exclusion is a complex concept that is relevant in terms of the health of vulnerable groups. Attempts have been made in the past to measure it, both at the population and the individual level. The aim of this research was to engage with a broad range of relevant stakeholders in Ireland in order to learn how they defined and conceptualised social exclusion. Semi-structured interviews were carried out with 24 participants selected using maximum variation sampling. One quarter of the interviewees were experts by experience. Participants included academic experts, the heads of organisations working nationally with socially excluded groups, politicians, clinicians, support workers and health service managers all with experience of working with socially excluded groups. The resulting definition of social exclusion was "the experience of lack of opportunity, or the inability to make use of available opportunities, thereby preventing full participation in society."From this, we developed a new model of the concept comprising three elements; Opportunities, Influencing factors and Social outcomes. Opportunities are the fundamental needs that are required to be met for a person to begin leaving social exclusion. Influencing factors are a mixture of the personal characteristics and more complex problems such as the intergenerational effects of disadvantage. Social outcomes include a person being accepted by wider society, and subsequently being able to participate. The conceptual framework we developed can contribute to a better understanding of the concept of social exclusion. The traditional policy focus on improving the needs of excluded people at the Opportunities level must continue, but must be complemented by tackling the problems at the levels of the Influencing factors and Social outcomes also. In terms of changes to practice, the measurement of the social exclusion status of people engaging with primary care and other services would be an important start in order to better understand the magnitude of the work required.</p

Obesity in adults: a 2022 adapted clinical practice guideline for Ireland

This Clinical Practice Guideline (CPG) for the management of obesity in adults in Ireland, adapted from the Canadian CPG, defines obesity as a complex chronic disease characterised by excess or dysfunctional adiposity that impairs health. The guideline reflects substantial advances in the understanding of the determinants, pathophysiology, assessment, and treatment of obesity. It shifts the focus of obesity management toward improving patient-centred health outcomes, functional outcomes, and social and economic participation, rather than weight loss alone. It gives recommendations for care that are underpinned by evidence-based principles of chronic disease management; validate patients' lived experiences; move beyond simplistic approaches of "eat less, move more" and address the root drivers of obesity. People living with obesity face substantial bias and stigma, which contribute to increased morbidity and mortality independent of body weight. Education is needed for all healthcare professionals in Ireland to address the gap in skills, increase knowledge of evidence-based practice, and eliminate bias and stigma in healthcare settings. We call for people living with obesity in Ireland to have access to evidence-informed care, including medical, medical nutrition therapy, physical activity and physical rehabilitation interventions, psychological interventions, pharmacotherapy, and bariatric surgery. This can be best achieved by resourcing and fully implementing the Model of Care for the Management of Adult Overweight and Obesity. To address health inequalities, we also call for the inclusion of obesity in the Structured Chronic Disease Management Programme and for pharmacotherapy reimbursement, to ensure equal access to treatment based on health-need rather than ability to pay

Myth or Memory? Recollections of Penal Times in Irish Folklore

Stories of priests being hunted down and murdered at Mass Rocks by priest catchers and soldiers during the Penal era in Ireland persist to the present day. Using Ó Ciosáin’s (2004) tripartite taxonomy of memory this paper explores the reasons why these images continue to dominate and reflect persecuted nature of Catholicism

The effects of interleukin-6 in a rat model of hepatocellular carcinoma

Interleukin-6 (IL-6) is integrally linked to normal hepatic growth and liver regeneration. The aim of the present study was to determine the expression and function of IL-6 signalling pathways m a rat model of hepatocellular carcinoma (HCC). In these studies HCC was charactensed by dysregulated expression of IL-6 signalling components in vivo and in vitro, including downregulated expression of the IL-6 receptor and increased IL-6 production and secretion. Recombinant human IL-6 (rhIL-6) treatment of isolated rat HCC cells and hepatocytes in vitro led to the activation of janus kinase-signal transducer and activator of transcription (Jak-STAT) and Ras-mitogen activated protein kinase intracellular signalling pathways Cyclin dependent kinase (cdk) inhibitor expression was increased m HCC cells after treatment with IL-6, an effect that was not observed in isolated hepatocytes. Decreases in cdk activity after rhIL-6 treatment result in a G0/G1 growth arrest in HCC cells. As a result of these in vitro studies, an algmate based cell microencapsulation system was developed to provide continual in vivo rhIL-6 delivery in a rat model of HCC CHO cells transfected to supra-express rhIL-6 were encapsulated in algmate and implanted in HCC tumour burdened rats resulting in high serum levels of biologically active rhIL-6 Recombinant human IL-6 was detected in both normal and tumour liver tissues resulting in activation of the Jak-STAT pathway. Due to tumour volume variability, no conclusions on the effects of rhIL-6 on tumour progression in vivo could be determined However, rhIL-6 treatment did result in increased total hepatic weight. Collectively this study demonstrates a role for IL-6 m HCC progression and identifies algmate encapsulated CHO cells as an effective method for rhIL-6 delivery in vivo which has potential in the study of normal and abnormal liver growth

Candida dubliniensis: An update

The increased incidence of fungal infections during the last decade has been well-documented. Given that one of the most important factors contributing to this phenomenon is the increased numbers of immunocompromised individuals, it is perhaps not surprising that species previously not associated with human disease and novel species previously unknown to science have been identified as potential pathogens (e.g., Penicillium marneffei , Emmonsia pasteuriana and Candida dubliniensis)

Identification and Expression of Multidrug Transporters Responsible for Fluconazole Resistance in Candida dubliniensis

Candida dubliniensis is a recently described Candida species associated with oral candidosis in human immunodeficiency virus (HIV)-infected and AIDS patients, from whom fluconazole-resistant clinical isolates have been previously recovered. Furthermore, derivatives exhibiting a stable fluconazole-resistant phenotype have been readily generated in vitro from fluconazole-susceptible isolates following exposure to the drug. In this study, fluconazole-resistant isolates accumulated up to 80% less [(3)H]fluconazole than susceptible isolates and also exhibited reduced susceptibility to the metabolic inhibitors 4-nitroquinoline-N-oxide and methotrexate. These findings suggested that C. dubliniensis may encode multidrug transporters similar to those encoded by the C. albicans MDR1, CDR1, and CDR2 genes (CaMDR1, CaCDR1, and CaCDR2, respectively). A C. dubliniensis homolog of CaMDR1, termed CdMDR1, was cloned; its nucleotide sequence was found to be 92% identical to the corresponding CaMDR1 sequence, while the predicted CdMDR1 protein was found to be 96% identical to the corresponding CaMDR1 protein. By PCR, C. dubliniensis was also found to encode homologs of CDR1 and CDR2, termed CdCDR1 and CdCDR2, respectively. Expression of CdMDR1 in a fluconazole-susceptible Δpdr5 null mutant of Saccharomyces cerevisiae conferred a fluconazole-resistant phenotype and resulted in a 75% decrease in accumulation of [(3)H]fluconazole. Northern analysis of fluconazole-susceptible and -resistant isolates of C. dubliniensis revealed that fluconazole resistance was associated with increased expression of CdMDR1 mRNA. In contrast, most studies showed that overexpression of CaCDR1 was associated with fluconazole resistance in C. albicans. Increased levels of the CdMdr1p protein were also detected in fluconazole-resistant isolates. Similar results were obtained with fluconazole-resistant derivatives of C. dubliniensis generated in vitro, some of which also exhibited increased levels of CdCDR1 mRNA and CdCdr1p protein. These results demonstrate that C. dubliniensis encodes multidrug transporters which mediate fluconazole resistance in clinical isolates and which can be rapidly mobilized, at least in vitro, on exposure to fluconazole